Development of sandwich Enzyme-Linked Immunosorbent Assay for the detection of porcine epidemic diarrhea virus in fecal samples

2018 ◽  
Vol 122 ◽  
pp. 151-155 ◽  
Author(s):  
Rui Gui ◽  
Hong-yan Shi ◽  
Wei Liu ◽  
Li Feng ◽  
Ke-li Yang ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Porcine Epidemic Diarrhea Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fecal Samples ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

Development and application of an indirect enzyme-linked immunosorbent assay using recombinant S1 for serological testing of porcine epidemic diarrhea virus

2019 ◽  
Vol 65 (5) ◽  
pp. 343-352
Author(s):  
Ying Shan ◽  
Yajie Liu ◽  
Ziqi Liu ◽  
Guowei Li ◽  
Cong Chen ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Porcine Epidemic Diarrhea Virus ◽  
Fluorescent Antibody ◽  
Area Under The Curve ◽  
Antibody Detection ◽  
Economic Losses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

Porcine epidemic diarrhea virus (PEDV) causes severe infectious diseases in all ages of swine and leads to serious economic losses. Serologic tests are widely accepted and used to detect anti-PEDV antibodies that could indicate PEDV infection or vaccination. In this study, PEDV recombinant S1 protein (rS1) was expressed with the Bac-to-Bac system and purified by nickel-affinity chromatography. An indirect enzyme-linked immunosorbent assay based on rS1 (rS1-ELISA) was then developed and optimized by checkerboard assays with serial dilutions of antigen and serum. Serum samples from 453 domestic pigs and 42 vaccinated pigs were analyzed by the indirect fluorescent antibody (IFA) test and rS1-ELISA. Taking IFA as a gold standard, rS1-ELISA produced a high sensitivity (90.7%) and specificity (94.6%) by a receiver operating characteristic (ROC) curve. In addition, ROC analysis also revealed that rS1-ELISA was consistent with IFA (area under the curve 0.9583 ± 0.0082). This rS1-ELISA was then applied to antibody detection in inactivated PEDV vaccinated pigs. The antibody could be detected 2–4 weeks after the first inoculation. These results indicated that the rS1-ELISA established in this study provides a promising and reliable tool for serologic detection of anti-PEDV IgG antibodies in infected or vaccinated pigs.

scholarly journals Immunohistochemical Detection of Porcine Epidemic Diarrhea Virus Compared to Other Methods

1998 ◽  
Vol 5 (3) ◽  
pp. 412-414 ◽  
Author(s):  
Franco Guscetti ◽  
Curzio Bernasconi ◽  
Kurt Tobler ◽  
Kristien Van Reeth ◽  
Andreas Pospischil ◽  
...  
Keyword(s):  
Porcine Epidemic Diarrhea Virus ◽  
Clinical Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Direct Immunofluorescence ◽  
Good Reliability ◽  
Diarrhea Virus ◽  
Pcr Method ◽  
Porcine Epidemic Diarrhea ◽  
Cryostat Sections ◽  
Formalin Fixed

ABSTRACT An immunohistochemistry method using formalin-fixed tissues, a direct immunofluorescence method using cryostat sections, an enzyme-linked immunosorbent assay (ELISA), and a PCR method were compared for diagnosis in a litter of weaned pigs that had been experimentally inoculated with wild-type porcine epidemic diarrhea virus (PEDV) and killed between 6 and 60 h after onset of diarrhea. The immunohistochemistry method proved to be as reliable as direct immunofluorescence for diagnosis of PEDV in tissues collected postmortem. The good reliability of ELISA for investigating clinical samples was confirmed, whereas the PCR method used was ineffective.

scholarly journals DEVELOPMENT AND BASIC FEATURES OF INDIRECT ENZYME-LINKED IMMUNOSORBENT ASSAY FOR DETECTION OF ANTIBODIES AGAINST PORCINE EPIDEMIC DIARRHEA

2018 ◽  
pp. 8-12
Author(s):  
A. V. Kanshina ◽  
A. S. Yakovleva ◽  
Ye. S. Orlova ◽  
A. V. Scherbakov
Keyword(s):  
Immunosorbent Assay ◽  
Disease Diagnosis ◽  
Test System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diarrhea Virus ◽  
Test Component ◽  
Disease Agent ◽  
Repeatability And Reproducibility ◽  
Porcine Epidemic Diarrhea ◽  
Basic Features

Spread of porcine epidemic diarrhea and its increased threat for the pig industry necessitate development of advanced techniques for the disease diagnosis. Use of recombinant antigens of the disease agent seems prospective. The recombinant antigen-based indirect enzyme-linked immunosorbent assay has been developed for the detection of antibodies against porcine epidemic diarrhea virus. The research performed allowed for determination of all necessary test conditions. Basic features of the developed method were determined during its validation. The test precision was above 90% pursuant to its repeatability and reproducibility, diagnostic specificity of the test amounted to 99.47%, and its sensitivity as compared to commercial ID Screen PEDV Indirect (IDvet, France) was 92% with very good compatibility of the two tests (k-criterion – 0.88) – 92%. The test-system demonstrates high stability upon the change of the key test component.

scholarly journals The N-terminal domain of Spike protein contributes to antigenicity difference between porcine epidemic diarrhea virus genotypes G1 and G2

2021 ◽  
Author(s):  
Qi-long Qiao ◽  
Ning Li ◽  
Ming-zhen Song ◽  
Jing Chen ◽  
Pan pan Yang ◽  
...  
Keyword(s):  
Amino Acid ◽  
Porcine Epidemic Diarrhea Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
S Protein ◽  
Diarrhea Virus ◽  
Terminal Domain ◽  
Porcine Epidemic Diarrhea ◽  
Amino Acid Mutations ◽  
Multiple Amino Acid ◽  
S Proteins

Porcine epidemic diarrhea virus (PEDV) strains have been clarified into two genotypes, G1 and G2, based on the sequence of the spike (S) gene. Amino acid mutations that distinguish the two PEDV genotypes were mostly located in the N-terminal domain (NTD) (aa 1-380) of S protein. The fact of increased outbreaks of G2 subtype PEDV and the failure of G1 subtype PEDV strain (CV777)-based vaccine in China since 2010 suggested that multiple amino acid mutations located in the NTD altered the antigenicity of S protein. To determine the role of the NTD of S protein in the antigenicity difference, the NTD of the CV777 vaccine strain (G1) and CH/ZMDZY/11 strain (G2) was expressed in E. coli, respectively. polyclonal antibodies (PAbs) against genotype-specific S proteins were prepared by immunizing BALB/c mice using purified S proteins. Antigenicity was systematically compared by detection of PAbs against two genotype PEDV strains and purified S proteins using Western blot, indirect enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), and serum cross-neutralization assay (SN). Consistent with the multiple amino acid mutations in the NTD of S protein, different antigenic cross-reactivity between the two genotypes was demonstrated. There was six-fold and more than twenty-fold difference in ELISA and SN titer between anti-CV777 S protein antibodies against G1 and G2 subtype strains, respectively. There was twofold and eight-fold difference in ELISA and SN titer between anti-ZMDZY S protein antibodies against G1 and G2 genotype strains, respectively. The results proved that the NTD of S protein contributes to the antigenicity difference between PEDV genotypes G1 and G2, and highlighted a G2 strain should be used to develop a vaccine for providing better protection against prevalent genotype of PEDV.

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